CN111298123A

CN111298123A - Antiviral composition and application thereof in resisting human influenza virus and avian influenza virus

Info

Publication number: CN111298123A
Application number: CN202010212067.7A
Authority: CN
Inventors: 朱寿会; 朱帅
Original assignee: Shenzhen Jinhuiqiu High Tech Co ltd
Current assignee: Shenzhen Jinhuiqiu High Tech Co ltd
Priority date: 2020-03-24
Filing date: 2020-03-24
Publication date: 2020-06-19

Abstract

The invention provides an antiviral composition and application thereof in resisting human influenza virus and avian influenza virus, belonging to the technical field of biological medicine. The antiviral composition consists of 7-110g of at least one detoxification, antibacterial and anti-inflammatory component, 5-60g of at least one qi-tonifying and blood-nourishing component and 3-85g of at least one kidney-tonifying and yang-supporting component, or consists of 7-110g of at least one antibacterial and anti-inflammatory component, 5-60g of at least one qi-tonifying and blood-nourishing component, 3-85g of at least one kidney-tonifying and yang-supporting component and 7-75g of at least one lung-ventilating and cough-relieving component. The antiviral composition provided by the invention can adapt to the variability of viruses through the synergistic effect of multiple components on the premise of no toxic or side effect, improves the effects of repairing the immune system of an infected person, resisting bacteria, diminishing inflammation, resisting organ inflammation of the infected person, and enhancing the antiviral effect, and has the effects of remarkably improving the cure rate and prolonging the survival rate.

Description

Antiviral composition and application thereof in resisting human influenza virus and avian influenza virus

Technical Field

The invention belongs to the technical field of biological medicines, and particularly relates to an antiviral composition and application thereof in resisting human influenza virus and avian influenza virus.

Background

Human diseases are caused by many biological factors, including bacteria, viruses, parasites, etc., and half of the diseases are caused by viruses, such as HIV, SARS virus, human (avian) influenza virus, hepatitis B, Ebola virus, papilloma virus of cervical cancer, etc., but viral diseases are difficult to treat, have fast spread and high mortality, and some viruses cause epidemic all over the world, such as novel coronavirus and influenza virus of this year.

However, because viruses have variability and recognition drug resistance, and western medicines and biological agents cannot adapt to the variation and recognition of the viruses, human beings have unprecedented difficulties in recognizing the viruses and searching for substances for killing the viruses. Chinese traditional medicine through a large amount of laboratory screening, found some Chinese herbal medicine and extracts with antiviral activity, and also demonstrated its antiviral effect in experimental animals, this contains rheum emodin of rhubarb source.

However, emodin is anthraquinone compound, which has poor solubility, is almost insoluble in water, is only soluble in alkali and some organic solvents, and has poor stability and is easy to be oxidized and deteriorated, so that patent applications CN106727482A and CN109528703A both structurally modify emodin, and although the modified emodin has greatly improved solubility, the stability is not improved therewith, and in the face of constantly mutated viruses, the virus inhibition effect which a single component can exert is very limited.

Disclosure of Invention

The invention provides an antiviral composition and application thereof in resisting human influenza virus and avian influenza virus, wherein the antiviral composition is a multi-component compound preparation, can adapt to virus variability through multi-component synergistic effect on the premise of no side effect, and has a remarkable inhibiting effect on the aspect of antivirus.

In order to achieve the aim, the invention provides a detoxifying and antiviral composition which comprises 7-110g of at least one antibacterial and anti-inflammatory component, 5-60g of at least one qi-tonifying and blood-nourishing component and 3-85g of at least one kidney-tonifying and yang-supporting component.

Preferably, the detoxifying, antibacterial and anti-inflammatory component is at least one selected from honeysuckle, agrimony, liquorice, trichosanthes root, isatis root, houttuynia cordata, forsythia suspensa and anthraquinone extract of rhubarb.

Preferably, the qi-tonifying and blood-nourishing component is at least one selected from fructus Jujubae, radix astragali, rhizoma Polygonati, Atractylodis rhizoma, radix Codonopsis and radix Angelicae sinensis.

Preferably, the kidney-tonifying yang-supporting component is at least one selected from the group consisting of epimedium, acanthopanax, yam, medlar, a temporary erection time, eucommia ulmoides, cistanche deserticola and bupleurum.

Preferably, the traditional Chinese medicine composition comprises 4-20 parts of cistanche, 3-30 parts of rhubarb anthraquinone extract, 5-35 parts of houttuynia cordata, 5-30 parts of isatis root, 2-20 parts of radix bupleuri, 5-30 parts of codonopsis pilosula, 3-25 parts of angelica sinensis and 1-15 parts of liquorice.

Preferably, the composition comprises 5-35 parts of rhizoma polygonati, 4-20 parts of eucommia ulmoides, 3-30 parts of rhubarb anthraquinone extract, 5-35 parts of honeysuckle, 1-15 parts of liquorice, 5-30 parts of trichosanthes root and 4-25 parts of barcan.

The invention provides an antiviral composition, which consists of 7-110g of at least one component for detoxifying, resisting bacteria and diminishing inflammation, 5-60g of at least one component for benefiting qi and nourishing blood, 3-85g of at least one component for tonifying kidney and supporting yang and 7-75g of at least one component for ventilating lung and relieving cough.

Preferably, the lung-ventilating and cough-relieving component is at least one selected from platycodon grandiflorum, adenophora stricta, fritillaria cirrhosa, rhodiola rosea and ophiopogon japonicus.

Preferably, the Chinese medicinal preparation comprises 3-30g of rhubarb anthraquinone extract, 10-60g of agrimony, 3-25g of rhodiola rosea, 10-30g of astragalus, 3-30g of herba epimedii, 5-30 g of radix ophiopogonis, 2-20 g of fritillaria and 5-30 g of fructus ziziphi chinensis.

Preferably, the medicine comprises 3-30 parts of rhubarb anthraquinone extract, 4-30 parts of fructus forsythiae, 2-20 parts of platycodon grandiflorum, 5-30 parts of acanthopanax, 10-40 parts of Chinese yam, 5-25 parts of medlar, 5-30 parts of adenophora stricta and 5-25 parts of bighead atractylodes rhizome.

Preferably, the anthraquinone extract of rhubarb comprises at least one of chrysophanol, emodin, physcion, aloe-emodin and rhein.

The invention also provides a preparation method of the antiviral composition according to the technical scheme, which comprises the following steps:

weighing the components according to the weight, removing impurities, cleaning, drying in the sun, and soaking in 8-20 times of purified water in a container for 0.5-3 days;

putting the soaking solution into a container, boiling with strong fire, keeping with slow fire for 30-600 min, filtering, and concentrating to obtain soft extract;

drying and pulverizing the soft extract, mixing with anthraquinone extract, adding appropriate amount of medicinal adjuvants, and making into pharmaceutically acceptable dosage forms.

Preferably, the dosage form can be capsule, tablet, granule, pill, oral liquid, etc.

The invention provides an application of the antiviral composition in resisting human influenza virus and avian influenza virus according to the technical scheme.

Preferably, the effective component of the antiviral composition is administered in an amount of 0.1 to 2 g/kg/d.

Compared with the prior art, the invention has the advantages and positive effects that:

the antiviral composition provided by the invention is a multi-component compound preparation, the safety of all raw materials in the preparation is recorded in Chinese pharmacopoeia of 2005 edition, the compound preparation is scientifically and reasonably matched, the compound preparation can adapt to the variability of viruses through the synergistic effect of multiple components on the premise of no side effect, and pharmacodynamics and toxicology experiments prove that the compound preparation not only has a remarkable inhibiting effect in the aspect of antivirus, but also has the effects of resisting bacteria and diminishing inflammation, improving immunity and shortening treatment course. Furthermore, the compound preparation can be prepared into various dosage forms such as tablets, capsules, pills, granules, pills, oral liquid and the like on the premise of simply changing the dosage form process, and can be conveniently used under various conditions.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The embodiment of the invention provides an antiviral composition, which consists of 7-110g of at least one component for detoxifying, resisting bacteria and diminishing inflammation, 5-60g of at least one component for tonifying qi and nourishing blood and 3-85g of at least one component for tonifying kidney and supporting yang.

Wherein the detoxifying, antibacterial and anti-inflammatory component is at least one selected from extracts of honeysuckle, agrimony, liquorice, trichosanthes root, isatis root, houttuynia cordata, forsythia and anthraquinone of rhubarb; the qi-tonifying and blood-nourishing component is at least one selected from fructus Jujubae, radix astragali, rhizoma Polygonati, Atractylodis rhizoma, radix Codonopsis and radix Angelicae sinensis; the kidney invigorating and yang supporting component is selected from at least one of herba Epimedii, radix Acanthopanacis Senticosi, rhizoma Dioscoreae, fructus Lycii, radix Paeoniae alba, Eucommiae cortex, Cistanchis herba and bupleuri radix.

The embodiment of the invention also provides an antiviral composition, which consists of 7-110g of at least one component for detoxifying, resisting bacteria and diminishing inflammation, 5-60g of at least one component for tonifying qi and nourishing blood, 3-85g of at least one component for tonifying kidney and supporting yang and 7-75g of at least one component for ventilating lung and relieving cough.

Wherein the detoxifying, antibacterial and anti-inflammatory component is at least one selected from extracts of honeysuckle, agrimony, liquorice, trichosanthes root, isatis root, houttuynia cordata, forsythia and anthraquinone of rhubarb; the qi-tonifying and blood-nourishing component is at least one selected from fructus Jujubae, radix astragali, rhizoma Polygonati, Atractylodis rhizoma, radix Codonopsis and radix Angelicae sinensis; the kidney invigorating and yang supporting component is selected from at least one of herba Epimedii, radix Acanthopanacis Senticosi, rhizoma Dioscoreae, fructus Lycii, radix Paeoniae alba, Eucommiae cortex, Cistanchis herba and bupleuri radix; the lung ventilating and cough relieving component is at least one of radix Platycodi, radix Adenophorae, Bulbus Fritillariae Cirrhosae, radix Rhodiolae and radix Ophiopogonis.

The raw materials in the formula have the following effects:

rhubarb, radix et rhizoma Rhei, is bitter, cold and nontoxic. It enters spleen, stomach, large intestine, liver and pericardium meridians. [ function and indications ] purgation and elimination of accumulation, clearing heat and purging fire, cooling blood and removing toxicity, removing blood stasis and dredging channels, promoting diuresis and eliminating jaundice. Can be used for treating constipation due to excessive heat accumulation, hematemesis, epistaxis, conjunctival congestion, pharyngeal swelling, carbuncle, furuncle, intestinal carbuncle, abdominal pain, blood stasis, amenorrhea, puerperal blood stasis, traumatic injury, damp-heat dysentery, jaundice, dark urine, stranguria, and edema; it can be used for external treatment of burn and scald. The wine rhubarb is good at clearing heat and toxicity in the upper jiao and blood system. It can be used for treating conjunctival congestion, pharyngeal swelling, and gingival swelling and pain. The cooked rhubarb has the functions of relieving the purgation, purging fire and removing toxicity. It can be used for treating pyocutaneous disease due to fire toxin. Rhubarb charcoal cools blood, removes stasis and stops bleeding. Can be used for treating blood heat with blood stasis and hemorrhage.

The agrimony (nature and taste) is smooth, pungent and astringent, cool and nontoxic, and enters heart and liver meridians. [ function and indication ] clear heat and promote diuresis, tonify deficiency and activate blood. It can be used for treating urinary tract infection, leucorrhea, dysentery, weakness after illness, sprain, and contusion.

Rhodiola root (nature and taste and meridian tropism) is flat, sweet, astringent and nontoxic, and enters lung and heart meridians. Has the functions and indications of invigorating vital energy, clearing away lung-heat, promoting intelligence, nourishing heart, arresting bleeding, dissipating blood stasis and eliminating swelling. Can be used for treating qi deficiency, asthenia, aversion to cold, short breath, asthenia, cough due to lung heat, hemoptysis, leucorrhea with diarrhea, and traumatic injury.

The astragalus root (nature and flavor and meridian tropism) is sweet in taste, slightly warm in nature and non-toxic; it enters spleen and lung meridians. [ Functions and indications ] tonify qi, strengthen superficies, expel toxin, expel pus, induce diuresis and promote granulation. Can be used for treating deficiency of vital energy, asthenia, chronic diarrhea, proctoptosis, spontaneous perspiration, edema, uterine prolapse, albuminuria due to chronic nephritis, diabetes, and unhealed wound.

Xianlingpi (nature, flavor and channel tropism) is pungent, warm, non-toxic, and enters liver and kidney channels. [ Functions and indications ] tonifying kidney yang, strengthening bones and muscles, dispelling wind-damp, relieving cough, dispelling phlegm, promoting hematopoiesis and immunity.

Acanthopanax root (nature, taste and channel tropism) is sweet, slightly bitter, warm and nontoxic; it enters spleen, lung, heart and kidney meridians. The composition has effects of invigorating qi, invigorating spleen, invigorating kidney, and tranquilizing mind, and can be used for treating spleen and lung qi deficiency, kidney deficiency, waist and knee pain, heart and spleen deficiency, insomnia, and amnesia.

The fructus Jujubae [ nature, flavor and meridian tropism ] is sweet, warm and nontoxic. It enters spleen and stomach meridians. [ Functions and indications ] tonify the middle-jiao and Qi, nourish blood and tranquilize the mind.

Radix Adenophorae (with nature, flavor and meridian tropism) is sweet, slightly bitter, slightly cold in nature and nontoxic. Meridian tropism: entering the lung; the stomach meridian. [ function and indication ] nourishing yin and clearing heat; moistening lung and eliminating phlegm; benefiting stomach and promoting fluid production. Chronic cough due to yin deficiency; cough with bloody phlegm; dry and cough with little sputum; deficiency-heat pharyngitis; thirst due to body fluid consumption.

Forsythia suspense (property, taste and channel tropism) is slightly cold, bitter in taste and non-toxic. The method comprises the following steps: lung, heart and small intestine. [ function and indication ] clearing away heat and toxic material, reducing swelling and resolving masses. Belonging to the category of heat-clearing herbs.

The root of balloonflower (nature, flavor and meridian tropism) is bitter, pungent, mild and nontoxic. The method comprises the following steps: the lung meridian. [ Functions and indications ] it is good at dispersing lung qi, eliminating phlegm and relieving sore throat, and also has the action of expelling pus, and is mainly indicated for cough with excessive phlegm, pharyngalgia hoarseness and lung abscess with pus discharge. Disperse lung qi, relieve sore throat, dispel phlegm, and expel pus.

The yam (nature, taste and channel tropism) is sweet, mild and nontoxic. Meridian tropism: it enters spleen, lung and kidney meridians. [ Functions and indications ] tonify spleen and stomach, promote the production of body fluid and benefit lung, tonify kidney and arrest seminal emission. Can be used for treating spleen deficiency, anorexia, chronic diarrhea, lung deficiency, cough, asthma, spermatorrhea, leukorrhagia, frequent micturition, and diabetes due to deficiency heat. The bran-parched rhizoma Dioscoreae has effects of invigorating spleen and invigorating stomach. Can be used for treating spleen deficiency, anorexia, diarrhea, loose stool, and leukorrhagia.

Medlar (nature, taste and meridian tropism) has mild nature, sweet taste and no toxicity. The method comprises the following steps: the liver and kidney meridians. [ Functions and indications ] nourishing liver and kidney, replenishing vital essence and improving eyesight. They belong to the yin tonics classified under the deficiency tonics.

Bighead atractylodes rhizome (with nature, taste and meridian tropism) is warm in nature, sweet in taste, bitter and nontoxic. The method comprises the following steps: spleen meridian and stomach meridian. [ function and indication ] invigorate the spleen, replenish qi, dry dampness and induce diuresis, stop sweating and prevent abortion. They belong to the category of qi tonics under deficiency-tonifying herbs.

Honeysuckle flower (nature and flavor with meridian tropism) is cold in nature and sweet in taste. The method comprises the following steps: lung, heart and stomach meridians. [ function and indication ] clearing away heat and toxic material, and cooling and dispelling wind-heat. Heat-clearing and toxicity-removing herbs belonging to the category of heat-clearing herbs

When used, the original flavor and meridian tropism are slightly warm in nature, sweet and pungent in flavor. The method comprises the following steps: kidney and liver meridians. [ Functions and indications ] tonify kidney yang, strengthen tendons and bones, dispel wind-damp. They belong to yang tonics classified under deficiency tonics.

Licorice root, radix Glycyrrhizae (nature and flavor and meridian tropism) is sweet and neutral in nature and enters heart, lung, spleen and stomach meridians. [ function and main treatment ] invigorating spleen and replenishing qi, eliminating phlegm and relieving cough, relieving spasm and pain, clearing heat and detoxicating, and harmonizing the drugs.

Trichosanthis radix (nature and flavor and meridian tropism) is sweet, slightly bitter and slightly cold. Meridian tropism: it enters lung and stomach meridians. [ function and indication ] clearing heat and promoting the production of body fluid, reducing swelling and discharging pus. Can be used for treating fever polydipsia, lung heat dry cough, internal heat diabetes, pyocutaneous disease and pyogenic infections.

Radix Isatidis (nature, taste and meridian tropism) is bitter and cold. Meridian tropism: the heart and stomach meridians. [ Functions and indications ] clear away heat and toxic material, cool blood and relieve sore throat.

Yu xing Cao (property and flavor and channel tropism) is pungent, slightly cold and nontoxic. Meridian tropism: it enters lung meridian. [ function and indication ] clearing away heat and toxic material, curing carbuncle and discharging pus, inducing diuresis and treating stranguria. Can be used for treating lung abscess with purulent vomiting, phlegm heat, cough and asthma, dysentery, pyretic stranguria, carbuncle, swelling, and sore.

Cistanche deserticola (nature, flavor and channel tropism) is sweet, salty, warm and nontoxic. Meridian tropism: it enters kidney and large intestine meridians. [ Functions and indications ] tonify kidney yang, replenish essence and blood, moisten intestines to relieve constipation. Can be used for treating sexual impotence, infertility, soreness of waist and knees, weakness of tendons and bones, constipation due to intestinal dryness.

Codonopsis pilosula (nature, flavor and channel tropism) is sweet, mild and nontoxic. Meridian tropism: it enters spleen and lung meridians. [ Functions and indications ] tonify the middle-jiao and qi, invigorate the spleen and benefit the lung. Can be used for treating spleen and lung deficiency, short breath, palpitation, anorexia, loose stool, asthma, cough, internal heat, and diabetes.

Dang Gui (property and flavor and meridian tropism) is sweet, pungent and warm. Meridian tropism: the liver, heart and spleen meridians. [ function and indication ] enriching blood and promoting blood circulation, regulating menstruation and relieving pain, and loosening bowel to relieve constipation. Can be used for treating blood deficiency, sallow complexion, giddiness, palpitation, menoxenia, amenorrhea, dysmenorrhea, asthenia cold, abdominal pain, constipation due to intestinal dryness, rheumatic arthralgia, traumatic injury, superficial infection, pyocutaneous disease.

Eucommia bark (nature, taste and meridian tropism) is sweet, warm and nontoxic. Meridian tropism: it enters liver and kidney meridians. [ Functions and indications ] tonify liver and kidney, strengthen tendons and bones, and prevent miscarriage. Can be used for treating lumbago due to kidney deficiency, weakness of bones and muscles, and pregnant bleeding and threatened abortion; hypertension is caused.

In the formula, the components for detoxifying, resisting bacteria and diminishing inflammation are monarch drugs, have the effects of detoxifying, resisting bacteria and diminishing inflammation, are the essential drugs for removing various toxins, the components for tonifying qi and nourishing blood and the components for tonifying kidney and supporting yang are ministerial drugs, are matched with the monarch drugs to play the effects of tonifying qi and strengthening exterior, expelling toxin and expelling pus, and can assist the body to promote the hematopoietic function and improve the immune function. The components are matched to play roles in clearing away heat and toxic materials, supporting toxicity and expelling toxin, promoting hematopoiesis and improving immunity. According to the severity of the patient, the composition can be matched with lung-ventilating and cough-relieving components to relieve the damage to the organism, and particularly, the protection to the lung can be strengthened. The feasible components listed in the components have similar medicinal effects, and only slight difference exists in the strength of the medicinal effect, so that doctors can reasonably match the components according to the actual conditions of patients. The raw materials of the provided antiviral composition are recorded in national standards, have considerable safety, and can prove that the effective components extracted based on the antiviral composition have theoretical basis for treating human influenza virus and avian influenza virus and have obvious inhibiting effect based on in vivo animal models and in vitro cell detection.

In the antiviral composition, the rhubarb anthraquinone extract comprises at least one of chrysophanol, emodin, physcion, aloe-emodin and rhein, and the rhubarb anthraquinone extract can be fluid extract of rhubarb anthraquinone, crushed body of dried rhubarb anthraquinone extract or further prepared particles by crushing.

In addition, the extraction raw material and the extraction method for the rhubarb anthraquinone compound are well known to the technicians in the field, and the principle is as follows: the anthraquinone glycoside is hydrolyzed into aglycone by dilute sulphuric acid solution, and extracted by hot chloroform/ether by utilizing the property that free anthraquinone can be dissolved in hot chloroform/ether. It is understood that there are many extraction methods for implementing the above principle, and the condition parameters can be adaptively adjusted according to different situations. In this regard, the extraction method is not within the scope of the present application, as long as the rhubarb anthraquinone-based extract or the particles thereof can be obtained.

Based on the above principle, the embodiment of the present application provides an existing extraction method, which specifically includes the following steps: 1) slicing radix et rhizoma Rhei, cutting, pulverizing to obtain radix et rhizoma Rhei coarse powder, and sieving with 80 mesh sieve to obtain radix et rhizoma Rhei fine powder; 2) collecting radix et rhizoma Rhei powder 20g, adding 20% H₂SO₄200ml, heating in water bath at 70 ℃ for 3-4 hours; 3) suction filtering, washing filter cake toDrying at about 70 deg.C after neutralization, and grinding into powder; 4) heating the extractant in the round-bottom flask to volatilize, leading the steam to go upwards through the gas-guide tube, cooling the steam into liquid in the condensing tube, and dripping the liquid into the tube provided with the sample filter paper cylinder to immerse the sample in the pure extractant; the substance to be extracted is gradually dissolved into the extracting agent from the sample; when the liquid level of the extracting agent in the sample tube gradually rises to be level with the upper end of the return tube, the extracted substances are carried to flow into the flask from the siphon tube on the side surface together under the siphon action, and then a round of circulation is completed; drying the filter cake, placing in a Soxhlet extractor, reflux-extracting with diethyl ether about 150ml for 3-4 hours to obtain diethyl ether extract, removing solvent, and volatilizing at room temperature to obtain rhubarb anthraquinone extract.

In order to more clearly and specifically introduce the antiviral composition provided by the embodiment of the invention and the application thereof in resisting human influenza virus and avian influenza virus, the invention takes the antiviral composition consisting of 3-30g of rhubarb anthraquinone extract, 10-60g of agrimony, 3-25g of rhodiola rosea, 10-30g of astragalus, 3-30g of epimedium, 5-30 g of dwarf lilyturf tuber, 2-20 of fritillary bulb and 5-30 of Chinese date as an example to carry out the following pharmaceutical pharmacodynamics experiment and toxicity experiment so as to show that the antiviral composition has no toxic or side effect, but has very obvious inhibiting effect on human influenza virus and avian influenza virus. It is understood that the pharmacological effects of other agents based on the inventive concept are not statistically different from the effects of the following experiments, and thus may be understood as the effects are the same or nearly similar, and thus the remaining experiments are not repeated.

1. Pharmacodynamic experiment

The pharmacodynamic experiment comprises an in vitro intracellular experiment and an in vivo animal model experiment.

1.1 intracellular assay

The intracellular assay is the first step in the evaluation of pharmacodynamics, i.e., the action of the drug on the monolayer of canine kidney passaged cells (MDCK), the ED is analyzed by comparing the study of the drug addition with the absence of drug, the MTT method and the hemagglutination method₅₀And T values, the results are shown in Table 1.

TABLE 1 drug dosage vs. drug efficacy Table

Note: "+ ++" indicates that more than 75% of the cells are diseased: "+ + + +" indicates 75% -50% of the cells were diseased; "+ +" indicates that 50% -20% of the cells were diseased; "+" indicates that less than 25% of the cells were diseased, "-" indicates that no cells were diseased.

Determination of ED by MTT and hemagglutination studies after treatment with SPASS115 software₅₀Are 1.27 μ gm and 1.08 μ gm, and TI values are 58.1 and 63.6.

1.2 animal model experiments

The animal model experiment is to study the drug effect of the drug in vivo, firstly, a human influenza virus (H1N1) model is made, and then an avian influenza (H5N1) model is made after the efficacy is proved.

The animal model comprises a control group and a treatment group, wherein the control group comprises a normal animal group and an infected animal group, the treatment group comprises a medicine group, a ribavirin group and a traditional Chinese medicine (Lianhua scourge) and the treatment group comprises a control before and after treatment and a control between the groups. The tested indexes include mortality, virus inhibition (by MT method and hemagglutination method), lung index, lung inhibition and virus detection rate. Through statistics of the indexes, the medicine has obvious inhibition effect on H5N1 and H1N1 and is better than ribavirin and traditional Chinese medicines (see tables 2, 3 and 4).

TABLE 2 Effect of drugs on pulmonary index of influenza Virus infected mice

TABLE 3 Effect of drugs on murine Lung tissue influenza Virus hemagglutination Titers

TABLE 4 Effect of drugs on the rate of detection of influenza Virus in mice (self-control)

In the pulmonary index and the inhibition rate thereof, the drug treatment groups have significant differences (P is less than 0.01) compared with the control group on the pulmonary virus hemagglutination titer and virus detection rate of mice (see Table 5).

TABLE 5 comparison of the effects of the three drugs

As shown in table 6, the results showed that the mortality of H1N1 in the drug group was 0 and that of the non-drug group was 10%; the mortality rate of the animals in the drug group of H5N1 is 0, and the mortality rate of the animals in the non-drug group is 30%. The detection rate of the virus of the H1N1 group before drug administration is 100 and the detection rate after drug administration is 18.75 by animal self control; the virus detection rate before the H5N1 group was 100, and 12.5 after the group was administered.

TABLE 6 self-control table for human and avian influenza models

Animal experiments show that the medicine has obvious inhibition effect on viruses in animals and no death case occurs in the medicine group. These indices were similar in either the H1N1 or H5N1 groups. The inhibition effect of the drug on the virus is calculated by statistics no matter in the H1N1 group or the H5N1 group, the inhibition rate of the drug group is higher than that of the non-drug group, and the significance is very remarkable (P is less than 0.01). These results are sufficient to demonstrate that the drug has significant inhibitory effect not only on HIN1, but also on H5 NI.

2. Toxicology studies

This example illustrates the safety of the antiviral compositions provided based on cytotoxicity, acute toxicity and chronic toxicity in three respects.

2.1 cytotoxicity

The cells affected in this example were the culture medium target cells TC740, and the drug concentration was 25mg/ml minimum and 1600mg/ml maximum, and the results showed that different drug doses were toxic to TC740, as the cell survival rate was inversely proportional to the drug concentration (see Table 7). TC of the drug by MTT assay detection and statistical calculation₅₀332.8 mg/ml.

TABLE 7 cytotoxic Effect of drug TC740

2.2 acute toxicity study

The animals used in this example were rats weighing between 100-120g, and half male and half female. The 72 white rats are randomly divided into 6 groups and 12 rats in each group, the 6 groups have the drug dosage of 17000mg, 25000mg, 35000mg, 50000mg, 70000mg and 105000mg per kilogram of body weight respectively, and the administration is performed once a day for 10 days. No death was observed in 25000mg/kg of animals, as confirmed by observation, and drug LD was calculated statistically₅₀It was 77964.3567 mg/kg.

2.3 Long term toxicity Studies

In this example, 101 rats with body weights between 110-120g, and 50 rats, 51 males and females, were randomly divided into 4 groups, three experimental groups (25 rats each) and one control group (26 rats). The drug doses are respectively as follows: the dosage of the large dose group is 0.92g/kg/d, the middle dose group is 0.31g/kg/d, the small dose group is 0.15g/kg/d, the dosages are 80 times, 40 times and 20 times of the dosage of the human, and the administration method is gastric lavage.

Experimental observations include animal performance, food intake, hair, fecal matter, blood indicators, blood biological indicators, mortality. The pathological examination includes heart, liver, stomach, kidney, lung, brain, testis, uterus, etc. The key point of pathological observation is the change of rat specimen and the change under the mirror. All data were statistically processed using the sps 10.0 software.

The results show that the rats do not die in 12 weeks after gastric lavage, the behaviors are normal, the feces are yellow brown, the properties are not abnormal, the indexes of urine in the experimental group are not obviously different from those of the control group (P >0.05), and the indexes of liver function in the experimental group are not obviously different from those of the control group (P > 0.05).

In terms of pathological anatomy, as shown in table 8, there was no significant difference in the body weight index of each organ between the experimental group and the control group. The difference between the pathological changes of the organs is not statistically significant. The animals in the experimental group had 16 lesions, and the animals in the control group had 5 lesions, the incidence of the former was 19.93%, the latter 19%, and there was no statistical difference between the two. The pathological nature is mostly mild cytopathic lesions, and also appears as pulmonary telangiectasia, alveolar septal thickening, bleeding spots in the brain and gliosis (only one animal), bleeding in the renal cortex, superficial inflammation in the stomach. The special lesion is liver cyst found in a rat. The above lesions are all mild pathological changes, while cellular degeneration is a reversible lesion. Therefore, the two groups of lesions are light in nature, and the number of lesion cases between the two groups of lesions is not obviously different. The incidence of organ disease in the two groups of animals is low, which indicates that the drug does not show long-term toxicity within safe dosage.

TABLE 8 incidence of lesions in organs of two groups of animals

As seen from the results in Table 8, the proportion of total lesions was within 20% in both the experimental group and the control group. The total abnormality rate of the experimental group was 19.93%, the control group was 19%, and there was no significant difference between the two. From the specific pathological nature, these lesions are all slight changes, some are reversible lesions (e.g. degeneration of liver cells), and we examined some animals before the experiment and found similar changes, which fully indicates that these changes should be independent of the drug intake. From the diseased organs, the probability of the occurrence of lung and stomach diseases is only slightly high (6.6%) but not more than 10% in the experimental group, and no disease occurs in heart, testis and ovary, which fully indicates that the long-term administration of the medicine has no influence on rats and does not damage the organs. The lesion ratio of the liver was 4% and 3.8% in the experimental group and the control group, respectively, and this ratio was low. The heaviest lesions in both groups of animals were granulomatous changes in hepatocytes in view of the nature of the lesions. Pathological granular pathological changes of the liver are reversible, the pathological changes are mild pathological changes, and ten normal rats are dissected before experiments to find similar pathological changes. The lesion proportion of the lung is 6.6%, the types of the lesions are mainly pulmonary interstitial telangiectasia and have very little interstitial bleeding, and the lesions are low in proportion firstly and light secondly. Lesions of the lungs are likely to be artificially created at the time of sacrifice. The pathological changes of the stomach are small in proportion, and the chronic superficial inflammation is presented, in fact, the stomach inflammation of the mouse is relatively common. Increased gastric irritation due to inadequate chewing of food tends to cause inflammation. Therefore, the lesions are not considered to be caused by the medicament, so that the medicament has safety to mice.

In conclusion, the antiviral composition provided by the invention has a very strong inhibitory effect on viruses such as H5N1 and H1N1, and has a broad spectrum. Moreover, the safety of all raw materials in the antiviral composition is recorded in Chinese pharmacopoeia 2005 edition, the collocation is scientific and reasonable, the medicine can adapt to the variability of viruses through the synergistic effect of multiple components on the premise of no side effect proved by toxicology, and pharmacodynamic experiments prove that the antiviral composition has a remarkable inhibiting effect on the aspect of antivirus. Furthermore, the antiviral composition can be prepared into various dosage forms such as tablets, capsules, oral liquid and the like on the premise of simply changing the dosage form process, and can be conveniently used under various conditions.

Claims

1. The antiviral composition is characterized by comprising 7-110g of at least one component for detoxifying, resisting bacteria and diminishing inflammation, 5-60g of at least one component for tonifying qi and nourishing blood and 3-85g of at least one component for tonifying kidney and supporting yang.

2. The antiviral composition as claimed in claim 1, wherein the detoxifying, antibacterial and anti-inflammatory component is at least one selected from the group consisting of extracts of honeysuckle flower, agrimony, licorice root, trichosanthes root, isatis root, houttuynia cordata, forsythia suspensa and anthraquinone.

3. The antiviral composition as claimed in claim 1, wherein the qi-invigorating and blood-nourishing component is at least one selected from fructus Jujubae, radix astragali, rhizoma Polygonati, rhizoma Atractylodis Macrocephalae, radix Codonopsis and radix Angelicae sinensis.

4. The antiviral composition as claimed in claim 1, wherein the kidney-tonifying and yang-supporting ingredient is at least one selected from the group consisting of epimedium, acanthopanax, yam, wolfberry, external-use plaster, eucommia, cistanche and bupleurum.

5. The antiviral composition according to any one of claims 1 to 4, wherein the composition comprises 4 to 20 parts of cistanche, 3 to 30 parts of rhubarb anthraquinone extract, 5 to 35 parts of houttuynia cordata, 5 to 30 parts of isatis root, 2 to 20 parts of radix bupleuri, 5 to 30 parts of codonopsis pilosula, 3 to 25 parts of angelica sinensis and 1 to 15 parts of liquorice.

6. The antiviral composition according to any one of claims 1 to 4, wherein the composition comprises 5 to 35 parts of Polygonatum sibiricum, 4 to 20 parts of eucommia ulmoides, 3 to 30 parts of anthraquinone extract of rhubarb, 5 to 35 parts of honeysuckle, 1 to 15 parts of licorice, 5 to 30 parts of Trichosanthis radix, and 4 to 25 parts of Bazaitian.

7. The antiviral composition according to claim 1, wherein the composition comprises 7-110g of at least one component for detoxification, antibiosis and anti-inflammation, 5-60g of at least one component for benefiting qi and nourishing blood, 3-85g of at least one component for tonifying kidney and supporting yang, and 7-75g of at least one component for ventilating lung and relieving cough.

8. The antiviral composition as claimed in claim 7, wherein the lung-ventilating and cough-relieving ingredient is at least one selected from the group consisting of Platycodon grandiflorum, Adenophora stricta, Fritillaria thunbergii, rhodiola rosea and Ophiopogon japonicus.

9. The antiviral composition according to claim 7 or 8, wherein the antiviral composition comprises 3-30g of anthraquinone extract of rhubarb, 10-60g of agrimony, 3-25g of rhodiola rosea, 10-30g of astragalus membranaceus, 3-30g of herba epimedii, 5-30 g of radix ophiopogonis, 2-20 g of fritillaria thunbergii and 5-30% of fructus ziziphi chinensis.

10. The antiviral composition according to claim 7 or 8, wherein the composition comprises 3-30 parts of rhubarb anthraquinone extract, 4-30 parts of forsythia, 2-20 parts of platycodon grandiflorum, 5-30 parts of acanthopanax senticosus, 10-40 parts of Chinese yam, 5-25 parts of medlar, 5-30 parts of adenophora stricta, and 5-25 parts of bighead atractylodes rhizome.

11. The antiviral composition according to claim 2, wherein the anthraquinone-based extract of rheum officinale comprises at least one of chrysophanol, emodin, physcion, aloe-emodin and rhein.

12. The method for preparing an antiviral composition according to claim 1, comprising the steps of:

13. The antiviral composition of claim 1, for use against human and avian influenza virus.

14. The use according to claim 13, wherein the antiviral composition is administered in an amount of 0.1 to 2g/kg/d as a dry product or in terms of a dry product.